Summary of Research Project Results Under the JSPS FY2000
"Research for the future Program"



1.Research Institution Tokyo Institute of Technology
 
2.Research Area Integrated Fields
 
3.Research Field Production of Novel Useful Substances through Integration of Life Sciences and Chemistry
 
4.Term of Project FY1996〜FY2000
 
5.Project Number 96I00302
 
6.Title of Project Mechanistic Analysis and Engineering of Secondary Metabolism Directed toward the Designer Enzyme Systems

7.Projetct Leader
Name Institution,Department Title of Position
KAKINUMA, Katsumi Tokyo Institute of Technology, Graduate School of Science and Engineering Professor

8.Core Members

Names Institution,Department Title of Position
EBIZUKA, Yutaka University of Tokyo, Graduate School of Pharmaceutical Sciences Professor
SAITO, Kazuki Chiba University, Faculty of Pharmaceutical Sciences Professor
ASANO, Yasuhisa Toyama Prefectural University, Faculty of Engineering Professor

9.Cooperating Researchers

Names Institution,Department Title of Position
EGUCHI, Tadashi Tokyo Institute of Technology, Graduate School of Science and Engineering Assoc. Prof.
FUJII, Isao University of Tokyo, Graduate School of Pharmaceutical Sciences Assoc. Prof.
KATO, Yasuo Toyama Prefectural University, Faculty of Engineering Assoc. Prof.

10.Summary of Research Results

Microbial and plant enzyme systems were studied to develop novel approaches for useful compounds.
The biosynthetic gene clusters of 2-deoxystreptamine-containing antibiotics, vicenistatin antitumor antibiotic and benzoisochromanequinone antibiotics were identified and sequenced. The mechanisms of key biosynthetic reactions were elucidated on the genetic, enzymic, and molecular levels. Analysis of genetic constitution of a series of PKSes gave new insight into the pathways and enzymes of polyketide biosyntheses. A carbocycle-forming biosynthetic enzyme was overproduced and applied to a novel chemo-enzymatic synthesis of catechol from sustainable glucose resource. Fungal PKS systems were also analyzed and a new polyketide biosynthetic pathway was elucidated. The triterpene synthase genes were cloned from various plants and sequenced. Construction of point-mutants and chimera poly-peptides made significant finding that the product selectivity of these synthases was controlled by rather small variation of amino acid and even a single mutation could alter the product contents. Differential display analysis of Perilla variants and genetic analysis allowed identifying the anthocyanin glucosyl transferase gene of the biosynthesis of anthocyanin pigments and the same gene was further identified from other plants as well. Cloning and over-expression of the responsible genes and chemical analysis precisely determined the mechanism and regulation of anthocyanidine formation in plants. Plant flavonoid biosynthetic enzymes were studied by genome analysis and a specific regulator gene was identified. In the metabolism of amino acids and related products, the genes for key biosynthetic enzymes of plant β-substituted alanines and microbial opine dehydrogenase were genetically cloned and overproduced. The latter enzyme and its site-specific mutants were structurally studied by X-ray crystallography and the reaction mechanism was precisely elucidated. Microbial aldoxime-nitrile pathway was discovered and aldoxime dehydratase was purified from soil bacteria. The molecular cloning of the responsible gene further allowed its overproduction and application for the enzymatic preparation of nitriles.

11.Key Words

(1)Enzyme Mechanism、(2)Molecular Analysis of Biosynthetic Genes、(3)Enzyme Engineering
(4)Engineered Biosynthesis、(5)Metabolic Control、(6)Antibiotics
(7)Amino Acid Metabolism、(8)Isoprenoids、(9)Alkaloids

12.References

[Reference Articles]
Author Title of Article
M. Kishida Diacetoneglucose Architechture as a Chirality Template. Part 9. Enantioselective Synthesis of (R)-Mevalolactone and (R)-[2H9]Mevalolactone on Carbohydrate Template.
Journal Volume Year Pages Concerned
J. Chem. Soc. Perkin I   1997 891-895

Author Title of Article
F. Kudo Kinetic Isotope Effect and Reaction Mechanism of 2-Deoxy-scyllo-inosose Synthase Derived from Butirosin-producing Bacillus circulans
Journal Volume Year Pages Concerned
J. Antibiot 50 1997 424-429

Author Title of Article
H. Morii A Novel Ether Core Lipid with an H-Shaped C80-Isoprenoid Hydrocarbon Chain from the Hyperthermophilic Methanogen Methanothermus fervidus
Journal Volume Year Pages Concerned
Biochem. Biophys. Acta 1390 1998 339-345

Author Title of Article
T. Eguchi Rational De Novo Design of NADH Mimic for Stereoselective Reduction Based on Molecular Orbital Calculation
Journal Volume Year Pages Concerned
Tetrahedron 54 1998 705-714

Author Title of Article
M. Morita Structural Diversity of Membrane Core Lipid of Extreme Halophile
Journal Volume Year Pages Concerned
Biosci. Biotechnol. Biochem 62 1998 596-598

Author Title of Article
T. Eguchi Multigram Synthesis of Mevalonolactone-d9 and Its Application to the Stereochemical Analysis of Saturation Reaction in the Biosynthesis of 2.3-Di-O-phytanyl-sn-glycerol Core of Archaeal Membrane Lipid as Studied by 1H-NMR
Journal Volume Year Pages Concerned
J. Am. Chem. Soc. 120 1998 5427-5433

Author Title of Article
Y. -F. Qiao Isolation and Characterization of A New Pyrano[4', 3':6, 7]naphtho[1, 2-b]xanthene Antibiotic FD-594
Journal Volume Year Pages Concerned
J. Antibiot. 51 1998 282-287

Author Title of Article
K. Kondo Structure and Biosynthesis of FD-594: A New Antitumor Antibiotic
Journal Volume Year Pages Concerned
J. Antibiot. 51 1998 288-295

Author Title of Article
H. Arai Absolute Stereochemistry of Vicenistatin, A Novel 20-Membered Macrocyclic Lactam Antitumor Antibiotic
Journal Volume Year Pages Concerned
Tetrahedron Lett. 39 1998 3181-3184

Author Title of Article
M. Otsuka Non-Fatty Acyl Polyketide Starter in the Biosynthesis of Vicenistatin, an Antitumor Macrolactam Antibiotic
Journal Volume Year Pages Concerned
Tetra hedron Lett. 39 1998 3185-3188

Author Title of Article
Y. Matsushima Enantioselective and Convergent Synthesis of the 20-Membered Lactam Aglycon of Vicenistatin Antitumor Antibiotic
Journal Volume Year Pages Concerned
J. Antibiot. 51 1998 688-691

Author Title of Article
E. Goetze Biosynthesis of Riboflavin. Stereochemistry of the 3, 4-Dihydroxy-2-butanone 4-phosphate Synthase Reaction
Journal Volume Year Pages Concerned
J. Org. Chem. 63 1998 6456-6457

Author Title of Article
N. Iwase Substrate Specificity of 2-Deoxy-scyllo-inosose Synthase. the Starter Enzyme in 2-Deoxystreptamine Biosynthesis, toward Deoxyglucose-6-phosphates and Proposed Mechanism
Journal Volume Year Pages Concerned
Biosci. Biotechnol. Biochem 62 1998 2396-2407

Author Title of Article
T. Eguchi Unique Solvent-dependent Atropisomerism of A Novel Cytotoxic Naphthoxanthene Antibiotic FD-594
Journal Volume Year Pages Concerned
J. Org. Chem. 64 1999 5371-5376

Author Title of Article
F. Kudo Purification and Characterization of 2-Deoxy-scyllo-inosose Synthase Derived from Bacillus circulans. A Crucial Carbocyclization Enzyme in the Biosynthesis of 2-Deoxystreptamine-containing Aminoglycoside Antibiotics
Journal Volume Year Pages Concerned
J. Antibiot. 52 1999 81-88

Author Title of Article
M. Ito A Novel Fungal Metabolite NG-061 Enhances and Mimics Neurotrophic Effect of Nerve Growth Factor (NGF) on Neurite Outgrowth in PC12 Cells
Journal Volume Year Pages Concerned
J. Antibiot. 52 1999 224-230

Author Title of Article
R. Bhandari Structure of NG-061, A Novel Potentiator of Nerve Growth Factor (NGF) Isolated from Penicillium miniolutem F-4647
Journal Volume Year Pages Concerned
J. Antibiot 52 1999 231-234

Author Title of Article
F. Kudo Molecular Cloning of the Gene for the Key Carbocycle-forming Enzyme in the Biosynthesis of 2-Deoxystreptamine-containing Aminocyclitol Antibiotics and Its Comparison witH Dehydroquinate Synthase
Journal Volume Year Pages Concerned
J. Antibiot. 52 1999 559-571

Author Title of Article
K. Kakinuma An Expeditious Chemo-Enzymatic Route from Glucose to Catechol by the Use of 2-Deoxy-scyllo-inosose Synthase
Journal Volume Year Pages Concerned
Tetrahedron Lett. 41 2000 1935-1938

Author Title of Article
T. Eguchi Unusual Double-Bond Migration as Plausible Key Reaction in the Biosynthesis of the Isoprenoidal Membrane Lipids of Methanogenic Archae
Journal Volume Year Pages Concerned
J. Chem. Soc. Chem. Commun   2000 1545-1546

Author Title of Article
T. Eguchi Synthesis of NG-061 and Its Analogs, and Their Biological Evaluation as an Enhancer of Nerve Growth Factor.
Journal Volume Year Pages Concerned
Chem. Pharm. Bull. 48 2000 1470-1473

Author Title of Article
Y. Ota Butirosin Biosynthetic Gene Cluster from Bacillus circulans.
Journal Volume Year Pages Concerned
J. Antibiot. 53 2000 1158-1167

Author Title of Article
M. Otsuka Biosynthetic Pathway of Macrolactam Polyketide Glycoside Antitumor Antibiotic Vicenistatins.
Journal Volume Year Pages Concerned
Tetrahedron 56 2000 8281-8286

Author Title of Article
K. Kakinuma New Approach to Multiply Deuterated Isoprenoids Using Triply Engineered Escherichia coli and Its Potential for Mechanistic Enzymology.
Journal Volume Year Pages Concerned
J. Am. Chem. Soc. 123 2001 1238-1239

Author Title of Article
Y. Matsushima Versatile Route to 2.6-Dideoxyamino Sugars from Non-sugar Materials: Syntheses of Vicenisamine and Kedarosamine
Journal Volume Year Pages Concerned
J. Chem. Soc. Perkin Trans 1   2001 569-577

Author Title of Article
Y. Matsushima Isolation and Structure Elucidation of Vicenistatin M. and Importance of the Vicenisamine Aminosugar for Exerting Cytotoxicity of Vicenistatin.
Journal Volume Year Pages Concerned
J. Antibiot. 54 2001 211-219

Author Title of Article
Y. Dekishima Preparation of Multiply Deuterated Carotenoids by Triply Engineered Escherichia coli with Perdeuterated Mevalonate.
Journal Volume Year Pages Concerned
Carotenoid Soience 4 2001 64-65

Author Title of Article
S. Sakuda Biosynthetic Studies on the Cyclopentane Ring Formation of Allosamizoline. an Aminocyclitol Derivative of the Chitinase Inhibitor. Allosamidin.
Journal Volume Year Pages Concerned
J. Org. Chem. (in press)   2001  

Author Title of Article
I. Fujii Cloning of the Polyketide Synthase Gene atX from Aspergillus terreus and Its Identification as the 6-Methylsalicylic Acid Synthase Gene by Heterologous Expression.
Journal Volume Year Pages Concerned
Mol. Gen. Genet 253 1996 1-10

Author Title of Article
T. Kushiro In vitro Conversion of 2.3-Oxidosqualene into Dammarenediol by Panax ginseng Microsomes
Journal Volume Year Pages Concerned
Biol. Pharm. Bull. 20 1997 292-294

Author Title of Article
M. Morita Molecular Cloning of Pea cDNA Encoding Cycloartenol Synthase and Its Functional Expression in Yeast
Journal Volume Year Pages Concerned
  20 1997 770-775

Author Title of Article
I. Fujii Anthracycline Biosynthesis in Streptomyces galilaeus
Journal Volume Year Pages Concerned
Chem. Review 97 1997 2511-2523

Author Title of Article
I. Fujii Structures and Functional Analyses of Fungal Polyketide Synthase Genes
Journal Volume Year Pages Concerned
Actinomycetol 12 1998 1-14

Author Title of Article
T. Kushiro β-Amyrin Synthase: Cloning of Oxidosqualene Cyclase that Catalyzes the Formation of the Most Popular Triterpene among Higher Plants
Journal Volume Year Pages Concerned
Eur. J. Biochem. 256 1998 238-244

Author Title of Article
A. Watanabe Product Identification of Polyketide Synthase Coded by Aspergillus nidulans wA Gene
Journal Volume Year Pages Concerned
Tetrahedron Lett. 39 1998 7733-7736

Author Title of Article
A. Watanabe Reidentification of Aspergillus nidulans wA Gene to Code for a Polyketide Synthase of Naphthopyrone
Journal Volume Year Pages Concerned
Tetrahedron Lett. 40 1999 91-94

Author Title of Article
K. Ichinose Proof that the actVI Genetic Region of Streptomyces coelicolor A3(2) is Involved in Stereospecific Pyran Ring Formation in the Biosynthesis of Actinorhodin
Journal Volume Year Pages Concerned
Bioorg. Med. Chem. Lett. 9 1999 395-400

Author Title of Article
K. Ichinose Biosynthetic Gene Cluster of Benzoisochromanequinone Antibiotics in Streptomyces spp. Identification of Genes Involved in Post-PKS Tailoring Steps
Journal Volume Year Pages Concerned
Actinomycetol. 12 1999 99-109

Author Title of Article
T. Kushiro Chimeric Triterpene Synthase. A Possible Model for Multifunctional Triterpene Synthase
Journal Volume Year Pages Concerned
J. Am. Chem. Soc. 121 1999 1208-1216

Author Title of Article
T. Kushiro Cryptic Regiospecificity in Deprotonation Step of Triterpene Biosynthesis Catalysed by New Members of Lupeol Synthase
Journal Volume Year Pages Concerned
Tetrahedron Lett. 40 1999 5553-5556

Author Title of Article
N. Funa A New Pathway for Polyketide Synthesis in Microorganisms
Journal Volume Year Pages Concerned
Nature 400 1999 897-899

Author Title of Article
I. Fujii Heterologous Expression and Product Identification of Colletotrichum lagenarium Polyketide Synthase Encoded by the PKS1 Gene Involved in Melanin Biosynthesis
Journal Volume Year Pages Concerned
Biosci. Biotech. Biochem. 63 1999 1445-1452

Author Title of Article
S. You Molecular Cloning and Sequencing of an Allium macrostemon cDNA Probably Encodimg Oxidosqualene Cyclase
Journal Volume Year Pages Concerned
Plant Biotechnology 16 1999 311-314

Author Title of Article
M. Shibuya Two Branches of the Lupeol Synthase Gene in the Molecular Evolution of Plant Oxidosqualene Cyclases
Journal Volume Year Pages Concerned
Eur. J. Biochem. 266 1999 302-307

Author Title of Article
H. Hayashi Molecular Cloning and Characterization of a cDNA for Glycyrrhiza glabra Cycloartenol Synthase
Journal Volume Year Pages Concerned
Biol. Pharm. Bull 23 2000 231-234

Author Title of Article
T. Taguchi Chemical Characterisation of Disruptants of the Streptomyces coelicolor A3(2) act VI Genes Involved in Actinorhodin Biosynthesis
Journal Volume Year Pages Concerned
J. Antibiotics 53 2000 144-153

Author Title of Article
M. Morita Molecular Cloning and Functional Expression of Triterpene Synthases from Pea (Pisum sativum); New α-Amyrin Producing Enzyme is a Multifunctional Triterpene Synthase
Journal Volume Year Pages Concerned
Eur. J. Biochem. 267 2000 3453-3460

Author Title of Article
T. Taguchi Identification of a Novel Shunt Product Produced by a Disruptant of the actVI-ORFA Gene Involved in the Biosynthesis of Actinorhodin in Streptomyces coelicolor A3(2)
Journal Volume Year Pages Concerned
Tetrahedion Lett. 41 2000 5253-5256

Author Title of Article
I. Fujii Enzymatic Synthesis of 1, 3, 6, 8-Tetrahydroxynaphthalene Solely from Malonyl Coenzyme A by a Fungal Iterative Type I Polyketide Synthase PKS1
Journal Volume Year Pages Concerned
Biochemistry 39 2000 8853-8858

Author Title of Article
T. Kushiro Mutational Studies on Triterpene Synthases. Engineering Lupeol Synthase into β-Amyrin Synthase
Journal Volume Year Pages Concerned
J. Am. Chem. Soc. 122 2000 6816-6824

Author Title of Article
T. Kushiro A Novel Multifunctional Triterpene Synthase from Arabidopsis thaliana
Journal Volume Year Pages Concerned
Tetrahedron Lett. 41 2000 7705-7710

Author Title of Article
A. Watanabe Aspergillus fumigatus alb1 Encodes Naphthopyrone Synthase When Expressed in Aspergillus oryzae
Journal Volume Year Pages Concerned
FEMS Microbiology Lett. 192 2000 39-44

Author Title of Article
I. Fujii Identification of Claisen Cyclase Domain in Fungal Polyketide Synthase WA, a Naphthopyrone Synthase of Aspergillus nidulans
Journal Volume Year Pages Concerned
Chemistry and Biology 8 2001 189-198

Author Title of Article
K. Ichinose Functional Complementation of Pyran Ring Formation in Actinorhodin Biosynthesis in Streptomyceces coelicolor A3(2) by Ketoreductase Genes from Granaticin Biosynthesis
Journal Volume Year Pages Concerned
J. Bacteriol (in press)   2001  

Author Title of Article
K. Saito Molecular cloning, characterization and expression of cDNA encoding phosphoserine aminotransferase involved in phosphorylated pathway of serine biosynthesis from spinach.
Journal Volume Year Pages Concerned
Plant Mol Biol. 33 1997 359-366

Author Title of Article
K. Saito Production of plant non-protein amino acids by recombinant enzymes of sequential biosynthetic reactions in bacteria.
Journal Volume Year Pages Concerned
Biol. Pharm. Bull. 20 1997 47-53

Author Title of Article
K. Saito Genomic structure and expression analyses of serine acetyltransferase gene in Citrullus vulgaris (watermelon).
Journal Volume Year Pages Concerned
Gene 189 1997 57-63

Author Title of Article
H. Takahashi Regulation of sulfur assimilation in higher plants: A sulfate transporter induced in sulfate-starved roots plays a central role in Arabidopsis thaliana.
Journal Volume Year Pages Concerned
Proa Natl. Acad. Sci. USA 94 1997 11102-11107

Author Title of Article
Z. Gong Cloning and molecular analysis of structural genes involved in anthocyanin biosynthesis and expressed in a forma-specific manner in Perilla frutescens.
Journal Volume Year Pages Concerned
Plant Mol Biol. 35 1997 915-927

Author Title of Article
M. Yamazaki Transformation of Perilla frutescens var. crispa using an Agrobacterium-Ri binary vector system.
Journal Volume Year Pages Concerned
Plant Biotechnology 14 1997 169-173

Author Title of Article
M. Kitajima Constituents of regenerated plants of Ophiorrhiza pumila; Formation of a new glycocamptothecin and predominant formation of (3R)-deoxypumiloside over (3S)-congener.
Journal Volume Year Pages Concerned
Tetrahedron Lett. 38 1997 8997-9000

Author Title of Article
O. Shirota Random amplified polymorphic DNA and restriction fragment length polymorphism analyses of Cannabis sativa.
Journal Volume Year Pages Concerned
Natural Med. 52 1998 160-166

Author Title of Article
A. Watanabe In vitro propagation. and restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) analyses of Angelica plants.
Journal Volume Year Pages Concerned
Plant Cell Rep 18 1998 187-192

Author Title of Article
T. Manabe Alliinase (S-alk(en)yl-L-cysteine sulfoxide lyase) from Allium tuberosum (Chinese chive): Purification, localization, cDNA cloning and heterologous functional expression.
Journal Volume Year Pages Concerned
Eur. J. Biochem. 257 1998 21-30

Author Title of Article
M. Noji Isoform-dependent differences in feedback regulation and subcellular localization of serine acetyltransferase involved in cysteine biosynthesis from Arabidopsis thaliana.
Journal Volume Year Pages Concerned
J. Biol. Chem. 273 1998 32739-32745

Author Title of Article
C. -L. Ho Molecular characterization of plastidic phosphoserine aminotransferase in serine biosynthesis from Arabidopsis.
Journal Volume Year Pages Concerned
Plant J. 16 1998 443-452

Author Title of Article
E. Yamagishi Molecular cloning of a cDNA encoding a novel UDP-glucose glucosyltransferase homologue from Arabidopsis thaliana (Accession No. ABO16819).
Journal Volume Year Pages Concerned
Plant Physiol 118 1998 1102

Author Title of Article
T. Nozaki Molecular cloning and characterization of the genes encoding two isoforms of cysteine synthase in the enteric protozoan parasite Entamoeba histolytica.
Journal Volume Year Pages Concerned
Mol. Biochem. Parasitol 97 1998 33-44

Author Title of Article
C. -L. Ho Regulation of serine biosynthesis in Arabidopsis: Crucial role of plastidic 3-phosphoglycerate dehydrogenase in non-photosynthetic tissues.
Journal Volume Year Pages Concerned
J. Biol. Chem. 274 1999 397-402

Author Title of Article
M. Yamazaki Molecular cloning and biochemical characterization of a novel anthocyanin 5-O-glucosyltransferase by mRNA differential display for plant formas regarding anthocyanin.
Journal Volume Year Pages Concerned
J. Biol. Chem 274 1999 7405-7411

Author Title of Article
K. Saito Direct evidence for anthocyanidin synthase as a 2-oxoglutarate-dependent oxygenase: molecular cloning and functional expression of cDNA from a red forma of Perilla frutescens.
Journal Volume Year Pages Concerned
Plant J. 17 1999 181-190

Author Title of Article
Y. Hatzfeld Identification of two putative nitrate transporters highly homologous to CHL1 from Arabidopsis thaliana (accession No. AJ011604 and AJ131464).
Journal Volume Year Pages Concerned
Plant Physiol 119 1999 805

Author Title of Article
C. -L. Ho Plastidic pathway of serine biosynthesis. Molecular cloning and expression of 3-phosphoserine phosphatase from Arabidopsis thaliana.
Journal Volume Year Pages Concerned
J. Biol. Chem. 274 1999 11007-11012

Author Title of Article
Z. Gong A light inducible Myb-like gene that is specifically expressed in red Perilla Frutescens and presumably acts as a determining factor of the anthocyanin forma.
Journal Volume Year Pages Concerned
Mol. Gen. Genet. 262 1999 65-72

Author Title of Article
H. Takagi Improved L-cysteine and L-cystine production by feedback inhibition-insensitive serine acetyltransferase from Arabidopsis thaliana in Escherichia coli.
Journal Volume Year Pages Concerned
FEMS Microbiol Lett. 179 1999 453-459

Author Title of Article
H. Takahashi Cloning of an Arabidopsis cDNA encoding a chloroplast localizing sulfate transporter isoform.
Journal Volume Year Pages Concerned
J. Exp. Botany 340 1999 1713-1714

Author Title of Article
Z. Gong A constitutively expressed Myc-like gene involved in anthocyanin biosynthesis from Perilla Frutescens: Molecular characterization. heterologous expression in transgenic plants and transactivation in yeast cells.
Journal Volume Year Pages Concerned
Plant Mol. Biol. 41 1999 33-44

Author Title of Article
H. Takahashi Identification of two leaf-specific sulfate transporters in Arabidopsis thaliana
Journal Volume Year Pages Concerned
Plant Physiol 121 1999 686

Author Title of Article
K. Inoue Determination of the sites required for the allosteric inhibition of serine acetyl-transferase by L-cysteine in plants.
Journal Volume Year Pages Concerned
Eur. J. Biochem. 260 1999 220-227

Author Title of Article
T. Leustek Sulfate transport and assimilation.
Journal Volume Year Pages Concerned
Plant Physiol 120 1999 637-643

Author Title of Article
K. Saito Regulation of sulfate transport and synthesis of sulfur-containing amino acids.
Journal Volume Year Pages Concerned
Curr. Opin. Plant. Biol   2000 188-195

Author Title of Article
Y. Hatzfeld Evidence for the existence of rhodanese (thiosulfate:cyanide sulfurtransferase) in plants: preliminary chracterization of two rhodanese cDNAs from Arabidopsis thaliana.
Journal Volume Year Pages Concerned
FEBS Lett. 470 2000 147-150

Author Title of Article
Y. Hatzfeld β-Cyanoalanine synthase is a mitochondrial cysteine synthase-like protein in spinach and Arabidopsis thaliana.
Journal Volume Year Pages Concerned
Plant Physiol 123 2000 1163-1171

Author Title of Article
Y. Hatzfeld Functional chracterization of a gene encoding a fourth ATP sulfurylase isoform from Arabidopsis thaliana.
Journal Volume Year Pages Concerned
Gene 248 2000 51-58

Author Title of Article
H. Takahashi The roles of three functional sulfate transporters involved in uptake and translocation of sulfate in Arabidopsis thaliana.
Journal Volume Year Pages Concerned
Plant J. 23 2000 171-182

Author Title of Article
M. Y. Hirai Biochemical and partial molecular characterization of bitter and sweet forms of Lupinus angustifolius, an experimental model for study of molecular regulation of qunolizidine alkaloid biosynthesis.
Journal Volume Year Pages Concerned
Chem. Pharm. Bull 48 2000 1458-1461

Author Title of Article
Y. Urano Molecular cloning and functional characterization of cDNAs encoding cysteine synthase and serine acetyltransferase that may be responsible for high cellular cysteine content in Allium tuberosum.
Journal Volume Year Pages Concerned
Gene 257 2000 269-277

Author Title of Article
Z. -Z. Gong Critical role of alanine-161 in Delila protein involved in regulation of anthocyanin pigmentation for transcriptional activation in yeast.
Journal Volume Year Pages Concerned
Plant Biotechnology 17 2000 331-335

Author Title of Article
K. Saito Feasible production of camptothecin by hairy root culture of Ophiorriza pumila.
Journal Volume Year Pages Concerned
Plant Cell Rep (in press)   2001  

Author Title of Article
W. Shitthithaworn Geranylgeranyl diphosphate synthase from Scoparia dulcis and Croton sublyratus. Plastid localization and conversion to a farnesyl diphosphate synthase by mutagenesis.
Journal Volume Year Pages Concerned
Chem. Pharm. Bull. 49 2001 197-202

Author Title of Article
M. Noji Cysteine synthase overexpression in Nicotiana tabacum confers tolerance to sulfurcontaining environmental pollutants.
Journal Volume Year Pages Concerned
Plant Physiol (in press)   2001  

Author Title of Article
C. -L. Ho Molecular biology of the plastidec phosphorylated serine biosynthetic pathway in Arabidopsis thaliana.
Journal Volume Year Pages Concerned
Amino Acids (in press)   2001  

Author Title of Article
J. Nakajima cDNA cloning and gene expression of anthocyanidin synthase from Torenia fournieri.
Journal Volume Year Pages Concerned
Plant Biotechnology 17 2000 331-335

Author Title of Article
Y. Asano Z-Phenylacetoaldoxime degradation by a novel aldoxime dehydratase from Bacillus sp. strain OxB-1
Journal Volume Year Pages Concerned
FEMS Micro 158 1998 185-190

Author Title of Article
K. L. Britton The Crystal Structure of the N-(1-D-Carboxyethyl)-L-norvaline dehydrogenase from Arthrobacter sp. strain 1C.
Journal Volume Year Pages Concerned
Nature Struct. Biol. 5 1998 593-601

Author Title of Article
Y. Kato Isolation and characterization of a bacterium possessing a novel aldoxime dehydratation activity and nitrile-degrading enzymes
Journal Volume Year Pages Concerned
Arch Microbiol. 170 1998 85-90

Author Title of Article
K. L. Britton Crystallization of Arthrobacter sp. Strain 1C opine dehydrogenase and its complex with NAD.
Journal Volume Year Pages Concerned
Acta Cryst. D. 5 1998 593-601

Author Title of Article
Y. Kato Nitrile hydratase involved in aldoxime metabolism from Rhodococcus sp. strain YH3-3 -purification and characterization-
Journal Volume Year Pages Concerned
Eur. J. Biochem. 263 1999 662-670

Author Title of Article
Y. Kato A new enzymatic method of nitrile synthesis by Rhodococcus sp.Strain YH3-3.
Journal Volume Year Pages Concerned
J. Molec. Catal. B:Enzymatic 6 1999 249-256

Author Title of Article
Y. Kato Opine dehydrogenase and secondary-amine dicarboxylic acids
Journal Volume Year Pages Concerned
Encyclopedia of Bioprocess Technology: Fermentation, Biocatalysis, and Bioseparation M. C. Flickinger and S. W. Drew (ed). John Wiley & Sons. Inc., New York   1999 1851-1858

Author Title of Article
Y. Kato A novel heme-containing lyase, phenylacetaldoxime dehydratase from Bacillus sp. strain OxB-1: purification. characterization. and molecular cloning of the gene.
Journal Volume Year Pages Concerned
Biochemistry 39 2000 800-809

Author Title of Article
Y. Kato Distribution of aldoxime dehydratase in Microorganisms.
Journal Volume Year Pages Concerned
Appl. Environ. Microbiol 66 2000 2290-2296

Author Title of Article
Y. Asano Selecton and directed evolution of new microbial biocatalysts and their application to organic synthesis.
Journal Volume Year Pages Concerned
Agric. Chem. Biotechnol. 43 2000 207-210

Author Title of Article
Y. Asano Overview of screening for new microbial catalysts and their uses in organic synthesis Selection and optimization of biocatalysts-
Journal Volume Year Pages Concerned
J. Biotechnol. (in press)      

Author Title of Article
K. Kakinuma Chirality Transfer on Carbohydrate Template: A Versatile Approach for Chiral Deuterium Labeling Svnthesis and Applications of Isotopically Labelled Compounds
Journal Volume Year Pages Concerned
    1998 543-546

Author Title of Article
H. Hayashi Molecular Cloning of Oxidosqualene Cyclase cDNA from Panax ginseng:The Isogene that Encodes β-Amyrin Synthase
Journal Volume Year Pages Concerned
Elsevier Science B. V. Amsterdam   1998 421-427

Author Title of Article
S. You Biosynthetic Studies on Steroid Saponins, Towards Natural Medicine Research in the 21st Century.
Journal Volume Year Pages Concerned
Elsevier Science B. V. Amsterdam   1998 429-441


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